Immunohistochemistry (Frozen sections) - PCNA antibody - Proliferation Marker (ab2426)

Ab2426 was used for immunohistochemistry, at a 1:500 dilution in hamster neural tissue, identified with a polyclonal secondary and DAB detection kit. Review by Kevin Bath submitted 18 June 2004

Immunocytochemistry/ Immunofluorescence - PCNA antibody - Proliferation Marker (ab2426)

ab2426 at 1/200 dilution staining rat skeletal muscle satellite stem cells by ICC/IF.

Cultured rat skeletal muscle satellite stem cells (SKMB) were 2% paraformaldehyde fixed for 15 minutes and then permealized with Triton-X100 prior to incubation with ab2426 overnight at 4°C. An Alexa-Fluor ® 488 conjugated donkey anti-rabbit antibody was used as the secondary.The image shows DAPI (blue-nuclear stain, upper left panel), PCNA (Green, upper right panel, showing nuclear localization in actively dividing cells), same SKMB cells -DIC (phase) image (lower left panel) and superimpose fluorescence image (lower right panel).

This image is courtesy of an Abreview submitted by Dr Mal Niladri

Western blot - PCNA antibody - Proliferation Marker (ab2426)

All lanes : Anti-PCNA antibody - Proliferation Marker (ab2426) at 1/200 dilution

Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

Lysates/proteins at 20 µg per lane.

Secondary
IRDye 680 conjugated Goat anti-rabbit IgG (H&L) at 1/15000 dilution

Predicted band size : 29 kDa
Observed band size : 29 kDa

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody - Proliferation Marker (ab2426)

ab2426 staining PCNA in murine skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using TRIS EDTA pH 8.2. Samples were then incubated with the primary antibody at a 1/2000 dilution for 1 hour at 37ºC. An undiluted HRP-conjugated rabbit polyclonal was used as secondary antibody.

Image courtesy of Laurence Sifre by Abreview.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody - Proliferation Marker (ab2426)

ab2426 staining PCNA in murine epithelial tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

Wound tissue sections (0.5 µm) were cut using a microtome and collected on slides. Sections were then de-waxed in xylene and rehydrated by successive immersion in descending concentrations of alcohol. The sections were then subjected for immunofluorescence staining. Briefly, tissue sections were blocked by incubated with 5% donkey serum for 1 hour and washed with phosphate-buffered saline (PBS). Sections were then incubated with ab2426 at a 1/500 for 1 hour at room temperature under humidified conditions. After primary antibody incubation, sections were washed with PBS and incubated with appropriate fluorescent secondary antibodies for 1 hour at room temperature. Sections were then washed with PBS, mounted with mounting medium containing DAPI.

Image from Liu S et al, J Cell Sci. 2010 Nov 1;123(Pt 21):3674-82. Epub 2010 Oct 12, Fig 3. DOI 10.1242/?jcs.070672

Western blot - Anti-PCNA antibody - Proliferation Marker (ab2426)

All lanes : Anti-PCNA antibody - Proliferation Marker (ab2426) at 1/300 dilution

Lane 1 : Whole tissue lysate prepared from mouse heart
Lane 2 : Whole tissue lysate prepared from mouse heart
Lane 3 : Whole tissue lysate prepared from mouse heart
Lane 4 : Whole tissue lysate prepared from mouse heart

Lysates/proteins at 50 µg per lane.

Secondary
HRP-conjugated pig anti-rabbit polyclonal at 1/3000 dilution
developed using the ECL technique

Predicted band size : 29 kDa
Observed band size : 29 kDa


Exposure time : 2 minutes

Image courtesy of an anonymous Abreview.